Bottom Line:
We further characterized the GUS expression patterns in the transgenic Arabidopsis lines.Our results show that both the 65 bp proximal promoter region and the 52 bp 5' UTR of AhLEC1B contain the key motifs required for the essential promoting activity.Moreover, AhLEC1B is preferentially expressed in the embryo and is co-regulated by binding of its upstream genes with both positive and negative corresponding cis-regulatory elements.

ABSTRACTLEAFY COTYLEDON1 (LEC1) is a B subunit of Nuclear Factor Y (NF-YB) transcription factor that mainly accumulates during embryo development. We cloned the 5' flanking regulatory sequence of AhLEC1B gene, a homolog of Arabidopsis LEC1, and analyzed its regulatory elements using online software. To identify the crucial regulatory region, we generated a series of GUS expression frameworks driven by different length promoters with 5' terminal and/or 3' terminal deletion. We further characterized the GUS expression patterns in the transgenic Arabidopsis lines. Our results show that both the 65 bp proximal promoter region and the 52 bp 5' UTR of AhLEC1B contain the key motifs required for the essential promoting activity. Moreover, AhLEC1B is preferentially expressed in the embryo and is co-regulated by binding of its upstream genes with both positive and negative corresponding cis-regulatory elements.

pone.0139213.g004: The sequence of 5′ flanking regulation region of peanut AhLEC1B gene and some major elements harbored in this region.The bold capital letter “A” represents the transcription start site (TSS), and other capital letters show different regulatory elements.

Mentions:
Based on the cDNA sequence of AhLEC1B, we further amplified the 5'UTR of the gene from the full-length cDNA library of Luhua14 developing seeds using nested 5' RACE. As a result, we obtained PCR products of about 400bp and 60bp (Fig 3). The transcription of AhLEC1B gene starts at the first ‘A’ within the sequence of CCAAACT. This sequence is located 83 bp upstream to the translation start codon ATG, and is consistent with the general feature in most eukaryotes (Fig 4).

pone.0139213.g004: The sequence of 5′ flanking regulation region of peanut AhLEC1B gene and some major elements harbored in this region.The bold capital letter “A” represents the transcription start site (TSS), and other capital letters show different regulatory elements.

Mentions:
Based on the cDNA sequence of AhLEC1B, we further amplified the 5'UTR of the gene from the full-length cDNA library of Luhua14 developing seeds using nested 5' RACE. As a result, we obtained PCR products of about 400bp and 60bp (Fig 3). The transcription of AhLEC1B gene starts at the first ‘A’ within the sequence of CCAAACT. This sequence is located 83 bp upstream to the translation start codon ATG, and is consistent with the general feature in most eukaryotes (Fig 4).

Bottom Line:
We further characterized the GUS expression patterns in the transgenic Arabidopsis lines.Our results show that both the 65 bp proximal promoter region and the 52 bp 5' UTR of AhLEC1B contain the key motifs required for the essential promoting activity.Moreover, AhLEC1B is preferentially expressed in the embryo and is co-regulated by binding of its upstream genes with both positive and negative corresponding cis-regulatory elements.

ABSTRACTLEAFY COTYLEDON1 (LEC1) is a B subunit of Nuclear Factor Y (NF-YB) transcription factor that mainly accumulates during embryo development. We cloned the 5' flanking regulatory sequence of AhLEC1B gene, a homolog of Arabidopsis LEC1, and analyzed its regulatory elements using online software. To identify the crucial regulatory region, we generated a series of GUS expression frameworks driven by different length promoters with 5' terminal and/or 3' terminal deletion. We further characterized the GUS expression patterns in the transgenic Arabidopsis lines. Our results show that both the 65 bp proximal promoter region and the 52 bp 5' UTR of AhLEC1B contain the key motifs required for the essential promoting activity. Moreover, AhLEC1B is preferentially expressed in the embryo and is co-regulated by binding of its upstream genes with both positive and negative corresponding cis-regulatory elements.